A5016604512- PI3K/AKT/mTOR signaling in cancer
- Hippo pathway signaling and YAP/TAZ
- RNA and protein synthesis mechanisms
- RNA Research and Splicing
- Protein Kinase Regulation and GTPase Signaling
- Polyamine Metabolism and Applications
- Cancer, Lipids, and Metabolism
- CRISPR and Genetic Engineering
- RNA modifications and cancer
- Metabolism, Diabetes, and Cancer
- Cancer, Hypoxia, and Metabolism
- Mast cells and histamine
- Coagulation, Bradykinin, Polyphosphates, and Angioedema
- Mitochondrial Function and Pathology
- Synthesis and Biological Activity
- Autophagy in Disease and Therapy
- Phosphodiesterase function and regulation
- Fungal Biology and Applications
- Genomics and Phylogenetic Studies
- Advanced Proteomics Techniques and Applications
- Bacterial Genetics and Biotechnology
- Molecular Biology Techniques and Applications
- Pancreatic function and diabetes
- Medical Imaging Techniques and Applications
- Endoplasmic Reticulum Stress and Disease
Yale University
2015-2024
University of New Haven
2015-2024
Massachusetts Institute of Technology
2006-2020
Whitehead Institute for Biomedical Research
2004-2016
Howard Hughes Medical Institute
2009-2016
Broad Institute
2006-2016
Allen Institute
2016
Harvard University Press
2013
Brigham and Women's Hospital
2013
Dana-Farber/Harvard Cancer Center
2013
The multiprotein mTORC1 protein kinase complex is the central component of a pathway that promotes growth in response to insulin, energy levels, and amino acids deregulated common cancers. We find Rag proteins—a family four related small guanosine triphosphatases (GTPases)—interact with an acid–sensitive manner are necessary for activation by acids. A mutant constitutively bound triphosphate interacted strongly mTORC1, its expression within cells made resistant acid deprivation. Conversely,...
The mammalian target of rapamycin (mTOR) kinase is the catalytic subunit two functionally distinct complexes, mTORC1 and mTORC2, that coordinately promote cell growth, proliferation, survival. Rapamycin a potent allosteric inhibitor with clinical applications as an immunosuppressant anti-cancer agent. Here we find Torin1, highly selective ATP-competitive mTOR directly inhibits both impairs growth proliferation to far greater degree than rapamycin. Surprisingly, these effects are independent...
Highly complex protein mixtures can be directly analyzed after proteolysis by liquid chromatography coupled with tandem mass spectrometry (LC−MS/MS). In this paper, we have utilized the combination of strong cation exchange (SCX) and reversed-phase (RP) to achieve two-dimensional separation prior MS/MS. One milligram whole yeast was proteolyzed separated SCX (2.1 mm i.d.) fraction collection every minute during an 80-min elution. Eighty fractions were reduced in volume then re-injected via...
Highlights•mTORC1 transcriptionally regulates amino acid transporters and enzymes via ATF4•mTORC1 controls ATF4 by regulating the translation stability of its mRNA•Control requires uORFs, but not changes in eIF2α phosphorylation•mTORC1 control instead employs 4E-BP repressorsSummaryThe mammalian target rapamycin complex 1 (mTORC1) is a master regulator cell growth that commonly deregulated human diseases. Here we find mTORC1 transcriptional program encoding metabolic through mechanism also...
Terminal oligopyrimidine (TOP) motifs are sequences at the 5' ends of mRNAs that link their translation to mTOR Complex 1 (mTORC1) nutrient-sensing signaling pathway. They commonly regarded as discrete elements reside on ∼100 mostly encode factors. However, full spectrum TOP and prevalence throughout transcriptome remain unclear, primarily because uncertainty over mechanism detects them. Here, we globally analyzed targets La-related protein (LARP1), an RNA-binding mTORC1 effector has been...
Abstract We have placed 7,600 cytogenetically defined landmarks on the draft sequence of human genome to help with characterization genes altered by gross chromosomal aberrations that cause disease. The are large-insert clones mapped chromosome bands fluorescence in situ hybridization. Each clone contains a tag is positioned genomic sequence. This genome-wide set sequence-anchored allows structural and functional analyses genome. resource represents first comprehensive integration...
Rapamycin is widely used as a complete inhibitor of the mTORC1 nutrient-sensitive signaling complex. Using novel ATP-competitive named Torin1, we have found that many functions regulate cap-dependent translation and autophagy are resistant to inhibition by rapamycin.
The mTOR protein is a master regulator of cell growth and proliferation, inhibitors its kinase activity have the potential to become new class anticancer drugs. Starting from quinoline 1, which was identified in biochemical assay, we developed tricyclic benzonaphthyridinone inhibitor 37 (Torin1), inhibited phosphorylation mTORC1 mTORC2 substrates cells at concentrations 2 10 nM, respectively. Moreover, Torin1 exhibits 1000-fold selectivity for over PI3K (EC(50) = 1800 nM) 100-fold binding...
To fully automate the sample introduction step for nanoscale microcapillary liquid chromatography−tandem mass spectrometry (LC−MS/MS) analyses, 75 μm i.d. × 14 cm capillary columns were interfaced with a commercial autosampler instrument using novel procedure which allowed dilute peptide samples to be transferred from AS loop injector column at flow rates up 5 μL min-1. On-column enrichment and desalting was demonstrated large volumes (>40 μL) by constructing vent 2 after entrance packed bed...
The mTOR mediated PI3K/AKT/mTOR signal transduction pathway has been demonstrated to play a key role in broad spectrum of cancers. Starting from the selective inhibitor 1 (Torin1), focused medicinal chemistry effort led discovery an improved 3 (Torin2), which possesses EC50 0.25 nM for inhibiting cellular activity. Compound exhibited 800-fold selectivity over PI3K (EC50: 200 nM) and 100-fold binding relative 440 other protein kinases. significantly bioavailability (54%), metabolic stability,...
Abstract mTOR is a highly conserved serine/threonine protein kinase that serves as central regulator of cell growth, survival, and autophagy. Deregulation the PI3K/Akt/mTOR signaling pathway occurs commonly in cancer numerous inhibitors targeting ATP-binding site these kinases are currently undergoing clinical evaluation. Here, we report characterization Torin2, second-generation ATP-competitive inhibitor potent selective for with superior pharmacokinetic profile to previous inhibitors....
Cell growth is a complex process shaped by extensive and coordinated changes in gene expression. Among these the tightly regulated translation of family growth-related mRNAs defined 5' terminal oligopyrimidine (TOP) motif. TOP mRNA partly controlled via eukaryotic initiation factor 4F (eIF4F), that recognizes cap structure. Recent studies have also implicated La-related protein 1 (LARP1), which competes with eIF4F for binding to ends. However, it has remained controversial whether LARP1...
An intensive recent effort to develop ATP-competitive mTOR inhibitors has resulted in several potent and selective molecules such as Torin1, PP242, KU63794, WYE354. These are being widely used pharmacological probes of mTOR-dependent biology. To determine the potency specificity these agents, we have undertaken a systematic kinome-wide profile their selectivity using chemical proteomics assays for enzymatic activity, protein binding, disruption cellular signaling. Enzymatic revealed that all...
Abstract Aberrant activation of the PI3K/mTOR pathway is a common feature many cancers and an attractive target for therapy, but resistance inevitably evolves as case any cancer cell–targeted therapy. In animal tumor models, chronic inhibition initially inhibits growth, over time, cells escape inhibition. this study, we identified context-dependent mechanism whereby upregulated proto-oncogene transcriptional regulators c-MYC YAP1. This was dependent on both constitutive ERK activity well...